We propose to establish a NIDA Neuroproteomics Research Center at the Yale University School of Medicine. More than 20 faculty members with an established history of studies of the molecular actions of psychostimulants and psychotropic drugs, as well as of other basic aspects of neurobiology, will work together with the W.M. Keck Foundation Biotechnology Resource Laboratory to create the Neuroproteomics Center. The main goal of the Center will be to apply high-throughput, state-of-the-art proteomic technologies to analyze adaptive changes in neuronal protein expression and regulation that occur in response to drugs of abuse. In addition, the Center will provide training in proteomics technologies, and will improve existing and develop new proteomies technologies that can be applied to biological questions related to the actions of drugs of abuse. The Center will include five cores: Protein Separation and Profiling, Protein Identification, Protein Microarray, Bioinformatics and Biostatistics, and Administration. A "lipidomics" component, included in the Protein Separation Core, will also allow the analysis of cellular lipids. The behavioral adaptations that accompany drug addiction are believed to result from both short and longterm adaptive changes in brain reward centers. To date, molecular studies of drugs of abuse have elucidated some of the transcriptional changes that occur in the addicted brain. However, little is known about the effects of drugs of abuse on the neuronal proteome. The Center will, through its highly interdisciplinary and collaborative organization, bring together Yale faculty with complementary expertise to gain a far deeper insight into how drugs of abuse alter expression and post-translational modification of proteins on a global scale. Methods that will be used will include MALDI-MS based biomarker analysis, two-dimensional chromatography, differential (fluorescence) gel electrophoresis (DIGE), isotope-coded affinity tag (ICAT), and antibody microarrays. Methods also will be developed for studying the brain phosphoproteome and these will be applied to studies of the actions of drugs of abuse. Specific goals of the research supported by the Center include analysis of the actions of opiates; the psychostimulants, cocaine and amphetamine; and nicotine on protein expression. Other studies will focus on the proteomic changes that occur both pre- and post-synaptically following the actions of drugs of abuse.